Scanning Tunneling Spectroscopic Studies of the Low-Energy Quasiparticle Excitations in Cuprate Superconductors

We report scanning tunneling spectroscopic (STS) studies of the low-energy quasiparticle excitations of cuprate superconductors as a function of magnetic field and doping level. Our studies suggest that the origin of the pseudogap (PG) is associated with competing orders (COs), and that the occurrence (absence) of PG above the superconducting (SC) transition T_c is associated with a CO energy Δ_(CO) larger (smaller) than the SC gap Δ_(SC). Moreover, the spatial homogeneity of Δ_(SC) and Δ_(CO) depends on the type of disorder in different cuprates: For optimally and under-doped YBa_2Cu_3O_(7−δ) (Y-123), we find that Δ_(SC) < Δ_(CO) and that both Δ_(SC) and Δ(CO) exhibit long-range spatial homogeneity, in contrast to the highly inhomogeneous STS in Bi_2Sr_2CaCu_2O_(8+x) (Bi-2212). We attribute this contrast to the stoichiometric cations and ordered apical oxygen in Y-123, which differs from the non-stoichiometric Bi-to-Sr ratio in Bi-2212 with disordered Sr and apical oxygen in the SrO planes. For Ca-doped Y-123, the substitution of Y by Ca contributes to excess holes and disorder in the CuO_2 planes, giving rise to increasing inhomogeneity, decreasing Δ_(SC) and Δ_(CO), and a suppressed vortex-solid phase. For electron-type cuprate Sr_(0.9)La_(0.1)CuO_2 (La-112), the homogeneous Δ_(SC) and Δ_(CO) distributions may be attributed to stoichiometric cations and the absence of apical oxygen, with Δ_(CO) < Δ_(SC) revealed only inside the vortex cores. Finally, the vortex-core radius (ξ_(halo)) in electron-type cuprates is comparable to the SC coherence length ξ_(SC), whereas ξ_(halo) ∼ 10ξ_(SC) in hole-type cuprates, suggesting that ξ_(halo) may be correlated with the CO strength. The vortex-state irreversibility line in the magnetic field versus temperature phase diagram also reveals doping dependence, indicating the relevance of competing orders to vortex pinning

Recent progress in the discovery of macrocyclic compounds as potential anti-infective therapeutics

Novel therapeutic strategies are urgently needed for the treatment of serious diseases caused by viral, bacterial and parasitic infections, because currently used drugs are facing the problem of rapidly emerging resistance. There is also an urgent need for agents that act on novel pathogen-specific targets, in order to expand the repertoire of possible therapies. The high throughput screening of diverse small molecule compound libraries has provided only a limited number of new lead series, and the number of compounds acting on novel targets is even smaller. Natural product screening has traditionally been very successful in the anti-infective area. Several successful drugs on the market as well as other compounds in clinical development are derived from natural products. Amongst these, many are macrocyclic compounds in the 1-2 kDa size range. This review will describe recent advances and novel drug discovery approaches in the anti-infective area, focusing on synthetic and natural macrocyclic compounds for which in vivo proof of concept has been established. The review will also highlight the Protein Epitope Mimetics (PEM) technology as a novel tool in the drug discovery process. Here the structures of naturally occurring antimicrobial and antiviral peptides and proteins are used as starting points to generate novel macrocyclic mimetics, which can be produced and optimized efficiently by combinatorial synthetic methods. Several recent examples highlight the great potential of the PEM approach in the discovery of new anti-infective agents

1003. Targeted Site-Specific Integration in Human Cells Using Designed Zinc Finger Nucleases

We have shown previously that human genome editing can be performed at high efficiency using designed zinc finger nucleases (ZFNs). ZFNs can be engineered to generate a double-strand break at a specific chromosomal location both in transformed and primary human cells. These breaks are repaired by the cell's own DNA repair machinery, and when a homologous extrachromosomal donor molecule is provided, a homology-directed repair process leads to highly efficient transfer of single-base-pair changes into the chromosome (Urnov et al. Nature 435: 646). ZFNs can be engineered to target virtually any chromosomal location (Pabo et al., Ann. Rev. Biochem. 70: 313), and function robustly in human hematopoietic stem cells, hence such localized gene modification may potentially be useful in the correction and treatment of certain monogenic diseases. Here we show that ZFNs can also be used for the preciseinsertion of a novel sequence into a pre-determined location in the human genome. Remarkably, this novel sequence can constitute a short patch sequence or several kilobases of one or more transgenes. We have observed ZFN-driven targeted integration into endogenous chromosomal loci in human cells of entire open reading frames and promoter-transcription units at a frequency of 5% in the absence of any selection. We also show the use of this process to disrupt with high efficiency an endogenous chromosomal locus with a selectable marker ORF. Finally, we describe our work on using ZFN-driven integration to insert a therapeutically-relevant transgene into a |[ldquo]|safe-harbor|[rdquo]| locus in the human genome, potentially avoiding the problem of insertional mutagenesis. Hence, the homology-directed repair process invoked by the ZFNs can be used to carry out high-efficiency targeted integration to potentially improve the safety and efficacy of gene therapy

Agonists and knockdown of estrogen receptor β differentially affect invasion of triple-negative breast cancer cells in vitro

Background: Estrogen receptor beta (ER beta) is expressed in the majority of invasive breast cancer cases, irrespective of their subtype, including triple-negative breast cancer (TNBC). Thus, ER beta might be a potential target for therapy of this challenging cancer type. In this in vitro study, we examined the role of ER beta in invasion of two triple-negative breast cancer cell lines. Methods: MDA-MB-231 and HS578T breast cancer cells were treated with the specific ER beta agonists ERB-041, WAY200070, Liquiritigenin and 3 beta-Adiol. Knockdown of ER beta expression was performed by means of siRNA transfection. Effects on cellular invasion were assessed in vitro by means of a modified Boyden chamber assay. Transcriptome analyses were performed using Affymetrix Human Gene 1.0 ST microarrays. Pathway and gene network analyses were performed by means of Genomatix and Ingenuity Pathway Analysis software. Results: Invasiveness of MBA-MB-231 and HS578T breast cancer cells decreased after treatment with ER beta agonists ERB-041 and WAY200070. Agonists Liquiritigenin and 3 beta-Adiol only reduced invasion of MDA-MB-231 cells. Knockdown of ER beta expression increased invasiveness of MDA-MB-231 cells about 3-fold. Transcriptome and pathway analyses revealed that ER beta knockdown led to activation of TGF beta signalling and induced expression of a network of genes with functions in extracellular matrix, tumor cell invasion and vitamin D3 metabolism. Conclusions: Our data suggest that ER beta suppresses invasiveness of triple-negative breast cancer cells in vitro. Whether ER beta agonists might be useful drugs in the treatment of triple-negative breast cancer, has to be evaluated in further animal and clinical studies

Comparative structural response of two steel bridges constructed 100 years apart

This paper presents a comparative numerical analysis of the structural behaviour and seismic performance of two existing steel bridges, the Infiernillo II Bridge and the Pinhao Bridge, one located in Mexico and the other in Portugal. The two bridges have similar general geometrical characteristics, but were constructed 100 years apart. Three-dimensional structural models of both bridges are developed and analysed for various load cases and several seismic conditions. The results of the comparative analysis between the two bridges are presented in terms of natural frequencies and corresponding vibration modes, maximum stresses in the structural elements and maximum displacements. The study is aimed at determining the influence of a 1 century period in material properties, transverse sections and expected behaviour of two quite similar bridges. In addition, the influence of the bearing conditions in the global response of the Pinhao Bridge was evaluated

Development of a single-chain, quasi-dimeric zinc-finger nuclease for the selective degradation of mutated human mitochondrial DNA

The selective degradation of mutated mitochondrial DNA (mtDNA) molecules is a potential strategy to re-populate cells with wild-type (wt) mtDNA molecules and thereby alleviate the defective mitochondrial function that underlies mtDNA diseases. Zinc finger nucleases (ZFNs), which are nucleases conjugated to a zinc-finger peptide (ZFP) engineered to bind a specific DNA sequence, could be useful for the selective degradation of particular mtDNA sequences. Typically, pairs of complementary ZFNs are used that heterodimerize on the target DNA sequence; however, conventional ZFNs were ineffective in our system. To overcome this, we created single-chain ZFNs by conjugating two FokI nuclease domains, connected by a flexible linker, to a ZFP with an N-terminal mitochondrial targeting sequence. Here we show that these ZFNs are efficiently transported into mitochondria in cells and bind mtDNA in a sequence-specific manner discriminating between two 12-bp long sequences that differ by a single base pair. Due to their selective binding they cleave dsDNA at predicted sites adjacent to the mutation. When expressed in heteroplasmic cells containing a mixture of mutated and wt mtDNA these ZFNs selectively degrade mutated mtDNA, thereby increasing the proportion of wt mtDNA molecules in the cell. Therefore, mitochondria-targeted single-chain ZFNs are a promising candidate approach for the treatment of mtDNA diseases

mTOR independent regulation of macroautophagy by Leucine Rich Repeat Kinase 2 via Beclin-1

Leucine rich repeat kinase 2 is a complex enzyme with both kinase and GTPase activities, closely linked to the pathogenesis of several human disorders including Parkinson’s disease, Crohn’s disease, leprosy and cancer. LRRK2 has been implicated in numerous cellular processes; however its physiological function remains unclear. Recent reports suggest that LRRK2 can act to regulate the cellular catabolic process of macroautophagy, although the precise mechanism whereby this occurs has not been identi ed. To investigate the signalling events through which LRRK2 acts to in uence macroautophagy, the mammalian target of rapamycin (mTOR)/Unc-51-like kinase 1 (ULK1) and Beclin-1/phosphatidylinositol 3-kinase (PI3K) pathways were evaluated in astrocytic cell models in the presence and absence of LRRK2 kinase inhibitors. Chemical inhibition of LRRK2 kinase activity resulted in the stimulation of macroautophagy in a non-canonical fashion, independent of mTOR and ULK1, but dependent upon the activation of Beclin 1-containing class III PI3-kinase